Method of lowering blood cholesterol level

ABSTRACT

The level of blood cholesterol in a living animal body in which a state of hypercholesterolemia exists is lowered by orally administering to said animal body a pharmaceutically effective amount of a polymer, which is (1) polymerized unsaturated carboxylic acid or anhydride, or (2) a copolymer of (a) an unsaturated monomer having, for Example, 2 to 30 carbon atoms and, (b) an unsaturated carboxylic acid, anhydride or derivative thereof as exemplified by octadecene-1/maleic anhydride copolymer. Polymers having lipophilic properties are a preferred class. A typical dosage is an amount which represents in the range of from about 0.01 to about 5.0% of the diet.

United States Patent 11 1 Fields et al. 1 Dec. 2, 1975 [5 METHOD OF LOWERING BLOOD 3.393168 7/1968 Johnson 260/297 CHOLESTEROL LEVEL 3,398,092 8/1968 Fields et al. 210/24 {75] Inventors: Joseph E. Fields, Ballwin; John H. OTHER IC T ONS Johnson Kilrkwoodboth of Cavallini et al., Potential Anticho/esterine-Mic Ann'- [73] Assignee: Monsanto Company, St. Louis, Mo. rheumatic Drugs 7/5/57 VOL P 3514-3517- Tennent et al., J. Lipid Research, 10/16, Vol. 5, pp. [22] F1led. Apr. 23, 1973 469L470 [21] Appl. No.: 353,832

Related Us Application Data Primary Exam1nerFreder1ck E. Waddell [63] Continuation-in-part of Ser. No. 188,577, Oct. 12,

1971, abandoned, which is a continuation-in-part of 1571 ABSTRACT 789,081,12111- 1969 abandoned- The level of blood cholesterol in a living animal body in which a state of hypercholesterolemia exists is low- [52] US. Cl. 424/78 ered by Orally administering to Said animal body a [51] A61K 27/00 pharmaceutically effective amount of a polymer, [58] Fleld of Search 188/557; 424/78 which is (i) polymerized unsaturated carboxyiic aCid or anhydride, or (2) a copolymer of (a) an unsatu- [56] References Clted rated monomer having, for Example, 2 to 30 carbon UNITED STATES PATENTS atoms and, (b) an unsaturated carboxylic acid, anhy- 2,378,629 6/1945 Hanford 260/78 dride Or derivative thereof as exemplified y ocmdec- 2,396,785 3/1946 l-lanford 260/78 aleic anhydride copolymer. Polymers having 3,157,595 11/1964 Johnson et a1... 210/78 lipophilic properties are a preferred class. A typical 2 12/1 N et 424/78 dosage is an amount which represents in the range of 3,308,020 3/1967 Wolf et al. 424/78 f about 001 to about of the diet 3,340,680 9/1967 Fields et al. /22 3,383,281 5/1968 Wolf et al. 424/78 9 Claims, No Drawings METHOD LOWERING BLOOD CHOLESTEROL LEVEL REFERENCE TO RELATED APPLICATIONS This application is a continuation-in-part of copending application Ser. No. 188,577, filed Oct. 12, 1971 and now abandoned, which application is in turn a continuation-in-part of copending application Ser. No. 789,081, filed Jan. 2, 1969, now abandoned.

BACKGROUND OF THE INVENTION This invention relates to a method for lowering the blood cholesterol level in a living animal body, in which a state of hypercholesterolemia exists. In a particular aspect this invention relates to a method for lowering the blood cholesterol level in a living animal body in which a state of hypercholesterolemia exists by orally administering to the living animal body a pharmaceutically effective amount of a polymer selected from the group consisting of (l) polymerized unsaturated carboxylic acid or anhydride and (2) a copolymer of (a) an unsaturated monomer having, for example, 2 to 30 carbon atoms, such as, alkene, alkylcarboxyalkene, phenylalkene and alkoxyalkene and, (b) an unsaturated carboxylic acid, anhydride or derivative thereof.

The link between high levels of blood cholesterol (hypercholesterolemia) and cardio-vascular disease in warmblooded vertebrates is well established. The malady is known not only to contribute to chronic conditions of high blood pressure, but to increase the risk of incapacitating and often fatal coronary attacks.

SUMMARY OF THE INVENTION It is an object of the present invention to provide a method for lowering the blood cholesterol in a living animal body in which a state of hypercholesterolemia exists.

It is a further object of the present invention to provide compositions which are effective in the aforesaid method in lowering blood cholesterol levels in a living animal body in which a state of hypercholesterolemia exists.

Other objects and advantages of the present invention will be apparent from the specification and appended claims.

The present invention resides in the discovery that the level of blood cholesterol in a living animal body in which a state of hypercholesterolemia exists is lowered by orally administering to said animal body a pharmaceutically effective amount of a polymer selected from the group consisting of (1 a polymerized unsaturated carboxylic acid, or anhydride (2) a copolymer of (a) an unsaturated monomer having, for example, 2 to 30 carbon atoms, and (b) an unsaturated carboxylic acid, anhydride or derivative thereof. By this method the blood cholesterol level in living animal bodies, including warm-blooded vertebrate animals, such as chickens, dogs, cats, cattle, swine and primates, for example monkeys, is effectively lowered.

DETAILED DESCRIPTION The polymer can be orally administered to the living animal body by any suitable means, and in any suitable form. For example, the polymer can be incorporated into ordinary foodstuffs and beverages containing nutritional values in anamount sufficient to produce the desired reduction of blood cholesterol. Also, the polymer can be incorporated into a pharmaceutical composition of the form customarily employed for oral administration. Pharmaceutical compositions containing the polymer may be in liquid form, for example, a solution or suspension specifically adapted for oral administration or in solid form, for example, a tablet, capsule, pill or packaged powder. Advantageously, the pharmaceutical composition containing the polymer can be prepared in unit dosage form using pharmaceutically acceptable carriers, such as, for example, starch, glucose, lactose, gelatin, sucrose, etc. and the like. If desired, the dosage unit can be made up in a sustained release form to give a controlled dosage over an extended period of time.

The amount or dosage of polymer administered to the living animal body will, of course, vary depending among other things, on the size of the living animal body, the particular living animal body to be treated, the level of blood cholesterol, and the general health of the living animal body, and any pharmaceutically effective amount may be employed. The dosage can be determined with regard to established medical practice. Generally the amount of polymer administered on a daily basis is in the range of from about 0.01 to about 5.0% of the total diet, and typically in the range of from about 0.05 to about 3.0%.

The polymer of use in the present invention may be water soluble or water-insoluble. Many of the normally water-soluble polymers are converted to the waterinsoluble form by introduction of sufficient crosslinks in the known manner. Crosslinking may be accomplished either during the preparation of the polymer or by subsequent treatment of the polymer to make the polymer insoluble in water. The water insolubility of the polymer can be varied by regulation of the degree of crosslinking of the polymer. The term water-insoluble as used herein is taken to mean that the polymer concerned does not dissolve in water or aqueous solutions, even though it may have such characteristics as a high degree of swelling, due to solvation by water even to the extent of existence in a gel form. Such characteristics are typically imparted by crosslinking.

As previously indicated, the polymers employed in the method of the present invention, are (l) polymerized unsaturated carboxylic acids or anhydrides, (2) copolymers of (a) an unsaturated monomer having, for example, 2 to 30 carbon atoms, and (b) an unsaturated carboxylic acid anhydride or unsaturated carboxylic acid derivative, preferably having a weight average molecular weight of at least 1,000 and a degree of polymerization of at least 8.

The polymer may advantageously be an EMA-type polymer.

Among the EMA-type polymers suitable for the practice of the instant invention are polymers and pharmaceutically acceptable salts of polymers having units of the formula wherein: R, and R are selected from the group consisting of hydrogen, halogen (preferably chlorine), alkyl of 1 to 4 carbon atoms (preferably methyl), cy-

ano, phenyl, or mixtures thereof; provided that not more than one of R and R is phenyl; Z is a bivalent radical (preferably alkylene, phenylalkylene, alkoxyalkylene, alkylcarboxyalkylene and aliphatic acyloxyalkylene) of l to 30 carbon atoms, q is zero or one, X and Y are selected from hydroxy, -O alkali metal, OR, OHNH OHR N, OHR NH, O- HRNl-l NRR, (Q),,-W(NRR), and (Q- ),,W(-OH) wherein x is l to 4 and p is zero or one, wherein R is selected from the group consisting of hydrogen, alkyl, phenylalkyl, or phenyl, in each case of l to 18 carbon atoms, wherein R is H or R, wherein Q is oxygen or NR'-, and wherein W is a bivalent radical preferably selected from alkylene, phenylene, alkylene amine and phenylalkylene having up to 20 carbon atoms, X and Y taken together can be oxygen or --NR, N-W-(NR'R') or -NW(N- R'R' R ).r wherein R, W, R have the meanings previously assigned and R" is alkyl of l to 18 carbon atoms, benzyl or aromatic-substituted benzyl. The units of the formula given above are recurring n being at least 8 and can be as much as 100,000 degrees of polymerization. When the units are recurring the symbols in the various recurring units do not necessarily stand for the same thing in all of the recurring units.

Many of these polymers suitable for the practice of the present invention or suitable after conversion to derivatives are commercially available.

The polycarboxylic acid polymers can be of the nonvicinal-type including those containing monomer units, such as acrylic acid, acrylic anhydride, methacrylic acid, crotonic acid or their respective derivatives, including partial salts, amides and esters or of the vicinal type, including maleic, itaconic, citraconic, a-dimethyl maleic, a-butyl maleic, a-phenyl maleic, fumaric, aconitic, a-chloromaleic, a-bromomaleic, a-cyanomaleic acids including their salts, amides and esters. Anhydrides of the foregoing acids are also advantageously employed.

Co-monomers suitable for use with the above polycarboxylic acid monomers include a-olefins, such as ethylene, Z-methyl-pentene-l, propylene, butylene, l-or 2-butene, l-hexene, l-octene, l-decene, l-dodecene, l-octadecene, and other vinyl monomers, such as styrene, a-methyl styrene, vinyltoluene, vinyl acetate, vinyl chloride, vinyl formate, vinyl alkyl ethers, e.g. methylvinylether, alkyl acrylates, alkyl methacrylates, acrylamides, methacrylamides, alkylmethacrylamides and alkylacrylamides, or mixtures of these monomers. Reactivity of some functional groups in the copolymers resulting from some of these monomers permits formation of other useful functional groups in the formed copolymer, including hydroxy, lactone, amine and lactam groups.

Any of the said carboxylic acids or derivatives, may be copolymerized with any of the other monomers described above, and any other monomer which forms a copolymer with carboxylic acids or derivatives. Although these copolymers can be prepared by direct polymerization of the various monomers, frequently they are more easily prepared by an after-reaction modification of an existing copolymer. Copolymers are :onveniently identified in terms of their monomeric :onstituents. The names so applied refer to the molecular structure and are not limited to the polymers preaared by the copolymerization of the specified mononers.

The initial copolymers of anhydrides and another monomer can be converted to carboxyl-containing copolymers by reaction with water, and to ammonium, alkali and alkaline earth metal and alkylamine salts thereof by reaction with alkali metal compounds, alkaline earth metal compounds, amines or ammonia. Other suitable derivatives of the above polymers include the alkyl or other esters and amides, alkyl amides, dialkyl amides, phenylalkyl amides or phenyl amides prepared by reacting carboxyl groups on the polymer chain with the selected amines or alkyl or phenylalkyl alcohol, as well as amino esters, amino amides, hydroxy amides and hydroxy esters, wherein the functional groups are separated by alkylene, phenyl, alkylene amine, alkylene oxide, phenylalkyl, phenylalkylphenyl, or alkylphenylalkyl or other aryl groups. Moieties bearing amine or amine salts including quaternary salt groups are conveniently formed by reaction of the carboxyls of their anhydride precursors, where applicable with polyfunctional amines such as dimethylaminopropylamine or dialkylaminoalcohols such as dimethylaminoethanol, the former forming an amide linkage with the polymer, or in certain cases at higher temperatures forming an imide linkage with vicinal carboxyls, and the latter forming an ester linkage. Such pendant free amine groups can then be converted, if desired, to their simple or quaternary salts.

Polymers of the above type include the following classes of polymers, and their derivatives: ethylene/- maleic anhydride copolymers, isobutylene/maleic anhydride copolymer, 2-methyl-pentene-l/maleic anhydride copolymers, styrene/maleic anhydride copolymers, vinylacetate/maleic anhydride copolymers, amethylstyrene/maleic anhydride copolymers, polymaleic anhydride polymers, polyacr'ylic anhydride polymers, polyacrylic acid polymers, octadecenel/maleic anhydride copolymers, loweralkylaminoloweralkylimide of octadecene-l/maleic anhydride copolymers, aliphatic ester of ethylene/maleic anhydride copolymers, vinylalkylether/ maleic anhydride copolymers, aliphatic methacrylate/methacrylamide copolymers, aliphatic methacrylate/diloweralkylaminoloweralkyl methacrylate copolymers, loweralkylaminoloweralkylimide of styrene maleic anhydride copolymers and polymethacrylic acid polymers.

Individual examples of such polymers include ethylene/maleic anhydride copolymer, the disodium salt of isobutylene/maleic anhydride copolymer, the calcium salt of styrene/maleic anhydride copolymer, hydrolyzed propylene/ maleic anhydride copolymer, the monopotassium salt of divinylether/maleic anhydride copolymer, hydrolyzed vinyl methyl ether/citraconic anhydride copolymer, half lauryl ester of octenel/maleic anhydride copolymer, octadecene-l/ maleic anhydride copolymer, ethylene/maleic acid copolymer, the dipotassium salt of isobutylene/maleic acid copolymer, the half amide half ammonium salt of isobutylene/maleic anhydride copolymer, ethylene/acrylic acid copolymer, ethylene/acrylic anhydride copolymer, half capryl ester of hexenel/acrylic anhydride copolymer, ethylene/aconitic anhydride copolymer, half ethylamide of styrene/maleic anhydride ethylene/fumaric acid copolymer, octylamide acid of ethylene/maleic anhydride copolymer, octadecylamide ammonium salt of vinylmethylether/maleic anhydride copolymer, dimethylaminopropylamide acid of divinylether/maleic anhydride copolymer, isobutylamide of vinyl acetate/maleic anhydride copolymer, me-

copolymer,

thiodide quaternary derivative of N, N-dimethylaminoethylamide of polymaleic anhydride, octadecyl ester ammonium salt of ethylene/itaconic anhydride copolymers, butylamine half amide of hexene-l/chloromaleic anhydride copolymer, the partial diamide of ethylene/- maleic anhydride copolymer, n-decylamide of decenel/maleic anhydride copolymer, N, N-diethylaminopropylamide ammonium salt of isobutylene/maleic anhydride copolymer, dimethyl sulfate quaternary salt of dimethylaminoethylamide of polymaleic anhydride, the partial half hexylamide of vinylmethylether/maleic an-- hydride copolymer, the diammonium salt of ethylene/- maleic anhydride copolymer, the monoamide acid of propylene/maleic anhydride copolymer, N-ethyl monoamide of divinylether/maleic anhydride copolymer, N- dodecyl monoamide of vinylmethylether/maleic anhydride copolymer, N, N-dimethylaminopropylimide of triacontene/maleic anhydride copolymer, N, N-dimethylaminopropyl monoamide of styrene/citraconic anhydride copolymer, n-butylmonoamide of polymaleic anhydride, N, N-diethylmonoamide ammonium salt of vinyl acetate/maleic anhydride copolymer, n-butylimide of ethylene/maleic anhydride copolymer, octadecylimide of polymaleic anhydride, N, N-dimethylaminopropylimide of styrene/maleic anhydride copolymer, dimethylsulfate quaternary salt of diethylaminopropylimide of divinylether/maleic anhydride copolymer, N, N-dimethylaminopropyl half amide of paramethyl styrene/maleic anhydride copolymer, methyliodide quaternary salt of dimethylaminohexyl half amide half ammonium salt of amethylstyrene/maleic anhydride copolymer, N, N-diethylaminoethyl half amide half sodium salt of isobutylene/maleic anhydride copolymer, partial lauryl ester of ethylene/maleic anhydride copolymer, vinyl octadecyl ether/maleic anhydride copolymer, stearyl methacrylate/ methacrylamide copolymer and stearyl methacrylate/N,N-dimethylaminoethyl methacrylate copolymer.

A particularly preferred class of polymers for use in the present invention are lipophilic polymers, that is, polymers which have a lipophilic grouping or groupings included therein. A lipophilic grouping or moiety typically contains 6 or more atom units and may be in any suitable form such as a polyalkylene or alkylene oxide containing 6 or more atom units or as an ester, amide or imide unit containing 6 or more atom units, for example, 6 to 30 carbon atoms formed by reaction of the carboxyl containing monomer with lipophilic amines or alcohols such as, for example, hexanol, octanol, octylamine, hexylamine, octadecanol, etc. and the like. Examples of such preferred polymers include the octadecylirnide of polymaleic anhydride, the methyl iodide quaternary salt of dimethylaminohexyl half amide half ammonium salt of a-methylstyrene/maleic anhydride copolymer, dodecyl monoamide of vinylmethylether/ maleic anhydride copolymer, the octadecyl ester ammonium salt of ethylene/itaconic anhydride copolymer, the decyla'mide of decene-l/maleic anhydride copolymer, .octadecene-1/ maleic anhydride copolymer etc. and the like.

Pharmaceutically acceptable alkaline earth metals and alkali metals, such as calcium, magnesium and potassium are useful in preparing conveniently administered forms of the polyelectrolyte polymers of this invention. The salts of metals such as magnesium, aluminum, zinc, iron, barium and bismuth are also useful in the present invention.

Representative EMA-type carboxylic acid or anhydride-olefin polymers, especially maleic acid or anhydrideolefin polymers of the foregoing type are known, for example, from US. Pat. Nos. 2,378,629;

r 2,396,785; 3,157,595; and 3,340,680. Generally, the

copolymers are prepared by reacting ethylene or other unsaturated monomer, or mixtures thereof, with the acid anhydride in the presence of a peroxide catalyst in an aliphatic or aromatic hydrocarbon solvent for the monomers but nonsolve nt for the interpolymer formed. Suitable solvents include benzene, toluene, xylene, chlorinated benzene and the like. While benzoyl peroxide is usually the preferred catalyst, other peroxides such as acetyl peroxide, butyryl peroxide, ditertiary butyl peroxide, lauroyl peroxide and the like, or any of the numerous azo catalysts, are satisfactory since they are soluble in organic solvents. The copolymer typically contains from about 25 to about (mole of the olefin and preferably contains substantially equimolar quantities of the olefin residue and the anhydride or acid residue; that is, a mole ratio of olefin to anhydride or acid in the range of from about 2:3 to about 3:2. Generally, the copolymer will have a degree of polymerization of 8 to 100,000 preferably about to 5,000, and a molecular weight of about 1,000 to 1,000,000, preferably about 10,000 to 500,000. The properties of the polymer, such as molecular weight, for example, are regulated by proper choice of the catalyst and control of one or more of the variables such as ratio of reactants, temperature, and catalyst concentration or the addition of regulating chain transfer agents, such as diisopropyl benzene, propionic acid, alkyl aldehydes, or the like. Numerous of these polymers are commercially available.

Derivatives containing basic or cationic groups can be prepared by any convenient procedure. Representative derivatives of polymers employed in the present invention are known to the art, for example, from US. Pat. No. 3,398,092. One group of useful derivatives are those in which the carboxyl groups are partially replaced with basic or cationic bearing moieties. For example, useful derivatives are conveniently formed by reaction of the carboxyls with polyfunctional amines such as dimethylaminopropylamine or dialkylaminoalcohols such as dimethylaminoethanol, the former forming an amide linkage with the polymer, or in certain cases at higher temperatures forming an imide linkage with the vicinal carboxyls and the latter forming an ester linkage. Such pendant free amine groups can then be converted, if desired, to their simple or quaternary salts.

Imides of a starting carboxyl or carboxylic acid anhydride containing polymer, e.g., EMA, are produced by:

A. Heating a limiting amount of a secondary or tertiary aminoloweralkylamine with the anhydride or carboxyl-containing form of the polymer in a suitable solvent (e.g. Xylene) at a temperature of about l40l50C. until water is no longer given off. Such a reaction simultaneously results in formation of imide groups in proportion to the amount of amine added and in the reformation of anhydride groups for the remainder of the polymer units. In this manner, imide-polymer products are formed which possess imide linkages, the remaining carboxyl groups, when present, being in the anhydride form.

B. Alternatively, a partial amidepolymer product may be converted to the partial imide polymer product by heating a partial amide-polymer product in vacuo at in the starting partial amide-polymer product.

Partial secondary or tertiary aminoloweralkylamides of the starting carboxyl or carboxylic acid anhydridecontaining polymer, e.g., EMA, are obtained by contacting the polymer with a limiting amount of the selected amine in suspension in a solvent such as benzene or hexane, resulting in formation of a partial amideacid-anhydride derivative of the polymer, or a corresponding amide-carboxylate derivative thereof. The number of amide groups is dependent upon the quantity of the amine used as compared with the quantity of polymer employed.

Partial aminoester-polymer products are most conveniently prepared by heating at reflux temperatures overnight a limiting quantity of the selected aminoalcohol and carboxyl or carboxylic acid anhydride containing polymer, e.g., EMA, in a dry organic solvent such as toluene or dimethylformamide and with the optional use of an acidic or basic catalyst such as p-toluenesulfonic acid or sodium alkoxide. The resulting product contains ester groups, carboxylic acid groups and anhydride groups, the respective numbers of which are determined by the quantity of aminoalcohol used in the reaction compared to the amount of polymer employed and, in some cases, by the temperature at which the reaction is carried out.

Suitable blocking and unblocking of the amine moiety of the reactant employed in preparing amides, esters or imides may be effected when required. Residual, nonmodified, polymer units may optionally be converted to neutral groups or units by attachment to the polymer molecule of compounds including alkylamines, aminoalcohols, and alcohols.

Alternatively, the cationic character of the polymer EXAMPLE 1 To determine the effect of polymers of the present invention in lowering blood cholesterol levels the following test was conducted.

A group of 144 Columbian male chicks (1 day old) was divided in 12 equal sub-groups. Each sub-group of 12 chicks was fed a basal diet and the supplement shown in Table 1 for a 3-week period. Throughout the test period water was supplied ad libitum. The basal diet employed was designed to raise the level of blood cholesterol in the chicks to which it was fed. On conclusion of the feeding period the chicks were sacrificed and plasma cholesterol level was determined by the general procedure described in Chin. Chim. Acta. 10 381-84 (1964) by Levine and Zak. Fecal cholesterol and fecal lipid were also determined. In the test the copolymers of diets 2-10 and 12 are linear and the copolymer of diet 11 is cross-linked. The results are set forth in Table 1:

The results of this test show that the polyelectrolyte polymers of the present invention are effective in low ering blood cholesterol and in increasing fat excretion.

TABLE 1 Plasma Fecal Level in Feed Cholesterol Cholesterol Fecal Lipid Diet Polymer by weight mg/ ml. dry weight dry weight 1. None (Control) 213 0.75 4.70 2. Octadecene-l/Maleic Anhydride Copolymer 0.5 183 1.06 6.9 3. Octadecene-llMaleic Anhydride Copolymer 1.0 150 1.32 9.28 4. Octadecene-l/Maleic Anhydride Copolymer 2.0 125 1.38 7.74 5. Octadecene-l/Maleic Anhydride Copolymer 3.0 107 1.43 10.29 6. N,N-Dimethylaminopropyl-imide of Octadecene-l/Maleic 3.0 114 1.58 24.7

Anhydride Copolymer 7. Partial lauryl ester of ethylene/Maleic Anhydride Copolymer 3.0 149 0.64 7 4 8. Vinyloctadecyl ether/Maleic Anhydride Copolymer 3.0 141 1.06 8.1 9. Stearyl methacrylate/Methacrylamide Copolymer 3.0 136 1.32 9.8 10. Stearyl methacrylate/N,N-dimethylamino ethyl methacrylate 3.0 1.56 22.4

copolymer 1 1. N.N-Dimethylaminopropylimide of styrene/maleic anhydride 3.0 107 1.53 20.3

copolymer l2. Octene-llmaleic Anhydride Copolymer 3.0 147 1.40 12.21

EXAMPLE 2 can be provided through incorporation of monomers which impart a basic or cationic character such as C- vinyl pyridines, vinyl amine, the several amino-substituted vinyl benzenes (or toluenes, etc. amine-bearing acrylates (or methacrylates, etc.), vinyl imioazole, etc.

The invention will be understood more fully by reference to the following specific examples. It is under- 65 stood that the examples are presented for the purpose of illustration only and are not intended as a limitation of the invention.

A group of 36 Vantress-Arbor Acre male chicks (1 60 day old) was divided into three equal subgroups. Each subgroup of 12 chicks was fed the Basal Diet of Exama 3 week period. Throughout the test period water was supplied ad libitum. On conclusion of the feeding period the chicks were sacrificed and plasma cholesterol level was determined by the procedure referred to in Example 1. 1n the test the copolymers of diet 14 and 15 were crosslinked. The results are set forth in Table 2.

The procedure of Example 2 was repeated. The polymers used and the results obtained are given in Table 3.

TABLE 3 PLASMA CHOLESTEROL DIET POLYMER mg/lOO ml 16 None 309 17 Ethylene/maleic anhydride (crosslinked) 275 18 Ethylene/maleic anhydride calcium salt (crosslinked) EXAMPLE 4 The procedure of Example 1 is repeated in all essential details with the exception that the polymer is polymaleic anhydride to lower the blood cholesterol level of the chicks.

EXAMPLE 5 The procedure of Example 1 is repeated in all essential details with the exception that the polymer is polyacrylic anhydride to lower the blood cholesterol level of the chicks.

EXAMPLE 6 The procedure of Example 1 is repeated in all essential details with the exception that the polymer is polyacrylic acid to lower the blood cholesterol of the chicks.

EXAMPLE 7 The procedure of Example 1 is repeated in all essential details with the exception that the polymer is polymethacrylic acid to lower the blood cholesterol level of the chicks.

EXAMPLE 8 Cattle having a high level of blood cholesterol are fed a daily ration containing from 0.01 to 5.0% of octadecene-l/maleic anhydride and the level of their blood cholesterol is lowered.

EXAMPLE 9 Heavy weight Hampshire hogs having a high level of blood cholesterol are fed a daily ration containing from 0.01 to 3.0% of N, N-dimethylaminopropylimide of octadecene-l/maleic anhydride copolymer and the level of their blood cholesterol is lowered.

EXAMPLE l0 Aged chimpanzees having a high level of blood cholesterol are fed a daily ration containing from 0.05 to 1.0% of octadecene- 1 /maleic anhydride copolymer and the level of their blood cholesterol is lowered.

EXAMPLE 1 1 This example shows the effectiveness of octadecenel/ maleic anhydride (substantially equimolar copolymer) in the method of the present invention. Twentyfour New Zealand white rabbits weighing an average of 2600 grams each were divided into three groups of equal number. The rabbits were fed ad libitum for 5 weeks a low cholesterol diet. The first group (control) was fed Purina Rabbit Chow brand rabbit feed supplemented with 2% animal tallow and 1% cellulose. The second group was fed the identical feed as the control with the exception that 1% octadecene-maleic anhydride was substituted for the 1% cellulose. The third group was fed the identical feed as the control with the exception that 1% CHOLESTYRAMINE, a crosslinked divinyl-benzene-polystyrene ion exchange resin containing quaternary ammonium groups was substituted for the 1% cellulose. On completion of the 5 week feeding period the animals were sacrificed and the following measurements were made: (1) plasma cholesterol, (2) plasma triglyceride, (3) liver cholesterol, and (4) liver fat. The measurements were averaged and are presented in Table 4. These results when considered as a whole indicate that higher a-olefinlmaleic anhydride copolymers are effective in lowering blood lipids, i.e., in the treatment of hypercholesterolemia.

EXAMPLE 12 This example shows the effectiveness of octadecenel/ maleic anhydride (substantially equimolar copolymer) in the method of the present invention. Forty New Zealand white rabbits weighing an average of 2600 grams each were divided into five groups of equal number. The groups were fed ad libitum for 5 weeks a high cholesterol diet. The first group (control) was fed Purina Rabbit Chow brand rabbit feed supplemented with 2% animal tallow, 1% cellulose and 0.2% cholesterol. The second group was fed the identical feed as the control with the exception that 0.5% octadecene-maleic anhydride was substituted for 0.5% of the cellulose. The third group was fed the identical feed as the control with the exception that 1% octadecene maleic anhydride was substituted for the 1% cellulose. The fourth group was fed the identical feed as the control with the exception that 0.5% CHOLESTYRAMINE was substituted for 0.5% of the cellulose. The fifth group was fed the identical feed as the control with the exception that 1% CHOLESTYRAMINE was substituted for the 1% cellulose. Feces were collected during the final day of the test. On completion of the 5-week feeding period the animals were sacrificed. The measurements performed in Example 11 above were carried out. Fecal fat was also measured. The measurements were averaged and are presented in Table 5.

EXAMPLE 13 This example shows the effectiveness of polymers useful in the present invention. Four hundred and sixtyeight l-day old chicks (Vantress-Arbor Acre Cockerels) were divided into 13 groups of equal number. Each group of chicks was fed ad libitum for 21 days a high cholesterol diet of soy bean meal, whole egg powder, minerals and vitamins. Cholesterol lowering additive in the amount indicated in Table 6 was added to the diet of groups 2-13. Group 1 served as a control. Feces were collected for a 24-hour period between days 19 1 1 and 20. On completion of the 21-day test the birds were sacrificed and the following measurements were made: (1) plasma cholesterol, (2) liver cholesterol, and (3) liver fat. The measurements were averaged and are 12 added to the diet of the third baboon. In each of the three animals the blood cholesterol level was found to be lower at the end of the eighth week than at the end of the third week with the blood cholesterol level at the presented in Table 6. 5 end of the eighth week being lower in the two animals fed the diet containing octadecene/maleic anhydride EXAMPLE l4 copolymer than in the animal fed the diet of CHOLES- This example shows the effectiveness of representa- TRYRAMINE. tive members of 1) a group of polymers which are co- Representative formulations embodying polymers polymers of higher a-olefins (-22 carbon atoms or 10 within the scope of the present invention are: more) and maleic acid or maleic anhydride and (2) a TABLET FORMULATION group of polymers which are copolymers of higher alkyl vinyl ethers (10-22 carbon atoms or more) and The following formulation provides for the manufacmaleic acid or maleic anhydride. The maleic acid 00- ture of 1,000 tablets: polymers were prepared by hydrolysis of the corresponding maleic anhydride copolymers in 5% acetic GRAMS acid in water at 50C for 24 hours followed by freeze 1 Octadecene-l/maleic anhydride copolymer 25 d i (2) Lactose g; Thirty-six female rabbits (1.5-2.0 kilograms each) fizg s ifs 2 were placed in individual cages. The rabbits were randomly divided into 12 groups (3 animals per group) for feeding purposes and fed ad libitum for 3 weeks high Thoroughly granulate a njnxture of 72 grams of Colin starch and the lactose with a paste prepared by discholesterol diets. The first group (control) was fed a solving 20 grams of cornstarch in 100 ml of hot distilled basal diet designed to induce hypercholesterolemla. t D th 1 ti n at 4O 45C and The died consisted of Purina Rabbit Chow brand rabbit e mg gran a O pass it through a No. l6-mesh screen. To the dried, feed supplemented with 2% animal tallow, 0.2% chod 1 ti dd a blended mixture of the lesterol and 1% cellulose. The other 11 groups were fed q l a on a trve ingredient (l) and the magnesium stearate. Thorthe identical basal diet as the control group with the exoughly blend and then press into tablets of 300 mg. ceptron that 1% of a copolymer useful in the present h invention was substituted for the 1% cellulose. On comeac pletion of the 3-week feeding period the animals were CAPSULE FORMULATION weighed and blood samples were taken. The samples I were analyzed for cholesterol (Technicon autoanat gi g i g i pmvldes for the manufaclyzer). At the beginning of the test, a blood sample was 0 p u e taken from each animal (heart puncture) and analyzed for cholesterol. All results are presented in Table 7. g I M The effectiveness of a tested compound is shown by U) g ggtgit mggff gggfi comparing the final plasma cholesterol of the animals copolymer 25 treated with the compound with the final plasma choh f gzz Stearate lesterol of the control animals. 40 g EXAMPLE 15 Mix active ingredient (l) with the lactose and blend Three baboons were fed a high cholesterol diet for 8 the magnesium stearate- Fill q gelatin Capsules weeks. At the beginning of the fourth week of feeding, Wlth 300 e e of the h e mlxhlre to Produce octadecene -1/maleic anhydride copolymer (0.5% by Capsules t s the aehve lhgredlehtweight of the total diet) was added to the diet of two of Whlle been desehhe'fl wlth refer the baboons and equal CHOLESTRYRAMINE, a ence to particular embodiments thereof, it W111 be apcrosslinked divinylbenzene-polystyrene ion exchange Pheelated that rhodlfieahohsehd vehahohs are Possible resin containing quaternary ammonium groups was Wlthout depamhg from the Invention- TABLE 4 PLASMA LIVER Cholesterol Triglyceride Cholesterol Fat *mg *mg of wet weight Group 1 (Control) 52 82 .38 6.89 Group 2 (1% Octadecene- 43 87 .36 5.12

maleic anhydride) Group 3 (1% 74 110 .39 5.69

CHOLESTYRAMINE) *mg per cc plasma.

TABLE 5 PLASMA LIVER FECES Cholesterol Triglyceride Cholesterol Fat Fat *mg *mg of wet weight of dry wt.

Group 1 (Control) 672 106 .92 i 7.80 4.51

Group 2 (0.5% octadecene-maleic 186 113 .47 5.98 6.03

anhydride) Group 3 (1.0% octadecene-maleic 89 56 .41 5.80 6.29

TABLE 5-continued PLASMA LIVER FECES Cholesterol Triglyceride Cholesterol Fat Fat *mg *mg 7: of wet weight of dry wt.

anhydride) Group 4 (0.5% CHOLESTYRAMlNE) 356 68 .79 7.36 7.12 Group 5 (1.0% CHOLESTYRAMlNE) 310 105 .74 7.18 6.04

*rng per 100 cc of plasma.

TABLE 6 Feed Plasma Liver Liver Excreta Exereta Weight Consumed Cholesterol Cholesterol Fat Cholesterol Fat Grams Grams mg 7c wet wt. wet wt. dry wt. dry wt.

Group 1 (control) 328 472 257 2.58 10.7 0.46 8.2 Group 2 (oocfittdecene-maleic anhydride) 360 483 250 1.26 8.9 0.97 6.

Group 3 (octidecene-maleic anhydride) 338 509 236 0.57 7.6 1.03 10.1

Group 4 (CHOLESTYRAMINE) (0.6%) 364 480 181 0.50 7.6 1.09 11.9 Group 5 (CHOLESTYRAMINE) (1.2%) 331 428 154 0.45 7.7 1.12 11.4 Group 6 (Octadeeene-maleic anhydride 354 491 226 0.89 8.2 0.83 11.2

dimethylaminopropylimide- 100% derivative) (0.6%)

Group 7 (Octadecene-maleic anhydride 315 456 258 dirnethylaminopropylimide- 67% derivative) (0.6%)

Group 13 (Stearyl methacrylate/ 369 455 274 0.8

methacrylic acid) (0.6%)

Group 8 (Stearylmethacrylate/dimethyl- 233 395 206 0.57 7.2 1.01 17.3

aminoethylmethacrylate) (0.6%)

Group 9 (Stearylmethacrylate/dimethyl- 370 501 262 0.80 14.1

aminoethylmethacrylate) (0.6%)

Group 10 (Vinyloctadecyl ether/ 361 463 242 0.87 7.9 1.1 1 12.0

maleic anhydride) (0.6%)

Group 11 (Vinyloetadecyl ether/ 360 514 298 0.97 9.5

maleic anhydride) (0.6%)

Group 12 (Stearyl methaerylate/ 358 439 282 0.73 9.5

methacrylic acid) (0.6%)

Group 13 (Stearyl methacrylate/ 369 455 274 0.84 9.5

methacrylic acid) (0.6%)

TABLE 7 Polymer Weight Gain in Grams For Plasma Cholesterol Plasma Cholesterol 3 Week Period (Average) lnitial mg.%* (Average) Final mg.7c* (Average) Group 1 (Control) 546 53 377 2 Octadecene/maleic 461 51 1 17 anhydride (2:3) (1) 3 Octadecene/maleic 487 123 84 acid (2:3) 4 Octadecene/maleic 436 147" anhydride (1:1) 5 Octadecene/maleic 288 61 1 l2 acid (1:1) 6 Decene/maleic 228 67 153 anhydride (1:1) 7 Decene/maleic acid 320 78 117 (1:1) 8 Octadecyl vinyl 460 91 162 ether/maleic anhy dride 9 Oetadecyl vinyl 279 64" 344 ether/maleic acid 10 Dodecyl vinyl 365 70 260 ether/maleic anhy- Y dride( l :1 l l Dodecyl vinyl 538 60 103 ether/maleic acid (1:1) 12 Hexene/maleic an- 670 60 472 hydride 1:1

*mg. pcr cc of plasma mole ratio of a-olefin to anhydride "*"1 animal died of diarrhea during test (average of 2 animals) 1 animal suffered severe weight loss due to diarrhea during test (average of 2 animals) "2 animals suffered severe weight loss due to diarrhea during test We claim: alkene of about 8 to about 22 carbon atoms and (2) 1. A method for lowering the blood cholesterol level maleic acid or anhydride, or a pharmaceutically acin a living animal in need thereof comprising orally adceptable amide, ester, imide, salt thereof or mixtures ministering to said animal an effective blood cholesthereof, said copolymer having a molecular weight in terol lowering amount of a copolymer of (1) an alpha the range of from about 1000 to 1,000,000.

6. The method of claim 1 in which the copolymer is a polyelectrolyte copolymer of an alkene of 10 to 22 carbon atoms.

7. The method of claim 1 in which the alkene has 10 to 18 carbon atoms.

8. The method of claim 1 in which the maleic component of the copolymer is a mixture.

9. The method of claim 1 in which the copolymer is a copolymer of maleic acid or anhydride. 

1. A METHOD FOR LOWERING THE BLOOD CHOLESTEROL LEVEL IN A LIVING ANIMAL IN NEED THEREOF COMPRISING ORALLY ADMINISTERING TO SAID ANIMAL AN EFFECTIVE BLOOD CHOLESTEROL LOWERING AMOUNT OF A COPOLYMER OF (1) AN ALPHA ALKENE OF ABOUT 8 TO ABOUT 22 CARBON TOMS AND (2) MALEIC ACID OR ANHYDRIDE, OR A PHARMACEUTICALLY ACCEPTABLE AMIDE, ESTER, IMIDE, SALT THEREOF OR MIXTURES THEREOF, SAID COPOLYMER HAVING A MOLECULAR WEIGHT IN THE RANGE OF FROM ABOUT 1000 TO 1,000,000.
 2. The method of claim 1 wherein the copolymer is octadecene-1/maleic anhydride.
 3. The method of claim 1 wherein the copolymer is deCene-1/maleic anhydride.
 4. The method of claim 1 wherein the copolymer is octadecene-1/maleic acid.
 5. The method of claim 1 in which the copolymer is a lipophilic copolymer of an alpha-alkene of 10 to 22 carbon atoms and maleic acid or anhydride.
 6. The method of claim 1 in which the copolymer is a polyelectrolyte copolymer of an alkene of 10 to 22 carbon atoms.
 7. The method of claim 1 in which the alkene has 10 to 18 carbon atoms.
 8. The method of claim 1 in which the maleic component of the copolymer is a mixture.
 9. The method of claim 1 in which the copolymer is a copolymer of maleic acid or anhydride. 